Imprint® Bisulfite DNA Modification

Sigma offers a complete solution for DNA methylation analysis with the Imprint® DNA Modification Kit. This product contains all of the reagents necessary for complete bisulfite conversion and subsequent purification of DNA samples. Only 50 picograms of DNA or 20 cells are needed for this procedure which can be completed in less than 2 hours.

Imprint® DNA Modification Kit

• Fastest Kit Available - procedure takes less than 2 hours to complete as compared to overnight methods
• Unparalleled Sensitivity - only 50 pg or 20 cells required for modification
• Complete Conversion - greater than 99% conversion of unmethylated cytosines
• Robust - more than 50% DNA recovery through treatment and purification

 View the Imprint® Methylated DNA Quantification Troubleshooting Questions

Figure 1. Bisulfite modification was performed on decreasing concentrations of human genomic DNA using Imprint® MOD50 and kits from two other suppliers. Methylation specific PCR (MSP) was then performed on all samples using β-Actin gene specific primers (109 bp amplicon). For all suppliers lanes from left to right are 100 ng, 10 ng, 1 ng, 100 pg, 50 pg and no sample respectively. Suppliers C and D (not shown) have no product at 50 pg. These results demonstrate the superior sensitivity of the Imprint® kit relative to other suppliers. Company Input DNA Time Required (hours) Imprint® >50 pg <2 Supplier A >100 pg <6 Supplier B >1 µg >24 Supplier C >1 ng >6 Supplier D >500 ng >10

Figure 2. 100 ng of human genomic DNA was bisulfite treated according to each supplier's protocol followed by qPCR analysis. The modified and unmodified primer sets within the β-Actin gene were then used to determine bisulfite yield. Ct values were compared to the Ct value of the 10 ng human genomic standard. Percent conversion was determined after bisulfite modification of 100 ng of human genomic DNA. Comparison of qPCR delta Ct values between modified and unmodified primers within β-Actin demonstrates conversion rates. The graph on the left highlights the superior yield and conversion rates of the Imprint® kit in relation to other suppliers.

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